Platelet-derived lysophosphatidic acid decreases endothelial permeability in vitro.
We previously reported that platelets release a soluble factor that decreases the solute permeability of cultured bovine aortic endothelial monolayers. This factor was characterized as heat stable, tryspsin sensitive, and not serotonin, adenosine, ADP, or ATP [F. R. Haselton and J. S. Alexander. Am. J. Physiol. 263 (Lung Cell Mol. Physiol. 7): L670-L678, 1992]. We now report its identity as lysophosphatidic acid (LPA). Endothelial permeability decreases rapidly, reversibly, and repeatedly when exposed to platelet supernatants. Continuous exposure produces a sustained decrease in permeability. Methanol extracts of platelet supernatants also decrease endothelial permeability. Treatment of methanol extracts of platelet supernatants with phospholipase B or alkaline phosphatase, which modify the structure of LPA, abolishes the permeability-decreasing activity. However, activity is unaffected by treatment with phospholipase A2. This pattern of enzyme inactivation is consistent with the structure of LPA. Furthermore, synthetic 1-oleoyl-LPA rapidly and significantly decreases endothelial permeability in a concentration-dependent manner. Platelet activation does not appear to be required to produce activity in supernatants from platelet isolations, since P-selectin expression is not increased and thromboxane B2 is < 14 pg/6,000 platelets. Our data show that platelets release a methanol-extractable compound with an enzyme degradation profile consistent with LPA, which decreases the permeability of endothelial monolayers in vitro. In vivo, LPA derived from platelets may be an important mediator of the transport barrier formed by the vascular endothelium.
Alexander
J S
JS
Vanderbilt University, Nashville, Tennessee 37235, USA.
Patton
W F
WF
Christman
B W
BW
Cuiper
L L
LL
Haselton
F R
FR
D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D001011:Aorta; D001792:Blood Platelets / Q000502:physiology*; D002417:Cattle; D002463:Cell Membrane Permeability / Q000187:drug effects / Q000502:physiology*; D002478:Cells, Cultured; D017077:Culture Media, Conditioned; D004730:Endothelium, Vascular / Q000187:drug effects / Q000502:physiology*; D005333:Fetus; D019793:Fluorescein; D006801:Humans; D007700:Kinetics; D008245:Lysophospholipase / Q000378:metabolism; D008246:Lysophospholipids / Q000097:blood* / Q000494:pharmacology; D010741:Phospholipases A / Q000378:metabolism; D054467:Phospholipases A2
D017077:Culture Media, Conditioned; D008246:Lysophospholipids; D010741:Phospholipases A; D054467:Phospholipases A2; D008245:Lysophospholipase; D000469:Alkaline Phosphatase; C032881:lysophosphatidic acid; D019793:Fluorescein